Background: Blood plasma/serum is a characteristic mixture of naturally occurring endogenous
fluorophores which are sensitive to endogenous and exogenous stress during physiological
as well as pathological processes in the body.
Methods: The structure of the patient's blood plasma/serum surfaces with critical limb ischemia and
healthy subjects were studied and compared using methods of synchronous fluorescence fingerprint
and atomic force microscopy, which are usually not used in clinical practice. The molecules of IGF-2,
HIF-1 and VEGF-A in the blood vessels of patients with a critical ischemic limb during the surgery
were analyzed by qRT-PCR and electrophoretic detection.
Results: Angiogenesis and also ischemia were detected in the ischemic blood vessels tissue of patients
as a significant increased expression of mRNA levels of HIF-1 and VEGF-A genes in comparison with
healthy subjects. The increased fluorescence intensity of proteins at wavelength λ = 280 nm/Δ60 nm
was observed in the blood plasma/serum of patients. The fluorescence spectroscopy and atomic force microscopy
revealed that the ischemic blood plasma and serum contains changed structures of proteins.
Conclusion: Spectroscopic signals can study ischemic changes and these can generally predict morphological
changes in the blood plasma/serum. Atomic force microscopy investigated structural
changes of proteins in the blood plasma/serum. Methods of molecular analysis detected significant hypoxia
in the blood tissue as significant increase of HIF-1 molecule and simultaneously significant angiogenesis
as a significant increase of VEGF-A molecules. New nontraditional techniques may contribute
to early diagnosis of the various vascular diseases in patients in future.
Keywords: Angiogenesis, atomic force microscopy, blood serum and plasma, critical limb ischemia, endogenous stress factors,
hypoxia, proteins, qRT-PCR, synchronous fluorescence spectroscopy.
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